Interest in producing large quantities of plasmid DNA has recently increased as a result of the rapid evolution of gene therapy and DNA vaccines. Hydrophobic chromatography is a popular technique in the downstream processing of plasmid DNA. However, the low capacity and the high mass transfer resistance of most commercially available packings for bio-macromolecules limit their application in a large-scale process. In this work, a hydrophobic absorbent with wide pores was synthesized by the solid porogenic method. Analyses by scanning electron microscopy and mercury intrusion porosimetry revealed that the matrix contained two families of pores, i.e., micropores smaller than 100 nm and superpores of 500-7300 nm. The superpores provided not only convective flow channels for the mobile phase, but also a large surface for biomolecules binding. So the chromatographic process can be operated at high flow rate with high column efficiency and low backpressure as identified on a 2-mL column (5mm i.d., 2cm length). With a loading up to 2.6 mg of 5.4 kb plasmid (pcDNA3) in 8mL feedstock and operated at a flow rate as high as 20 cm/min, nearly 100% of plasmid was recovered with a purity of 100%. The results indicate that the hydrophobic medium is promising for high-speed purification of plasmid DNA.

研究了多种添加剂促进变性还原溶菌酶复性的作用。考察了添加剂浓度及变性剂盐酸胍浓度对复性收率的影响。结果表明，精氨酸、乙酰胺、丙酮、硫脲及甘油均能有效促进变性溶菌酶复性，并且存在最佳的添加剂浓度使变性溶菌酶的复性收率最大。在促进复性中，乙酰胺等结构类似物与盐酸胍具有相同作用，因此，在降低复性液中盐酸胍浓度的同时，适当提高乙酰胺浓度即可获得较高的复性收率。当盐酸胍浓度为0.2mol/L时，复性收率达到90%时的乙酰胺浓度为2mol/L，但降低盐酸胍浓度至0.06mol/L时，达到相同变性收率的乙酰胺浓度需要4mol/L。甘油与盐酸胍存在着协同作用，在一定浓度的盐酸胍存在下，添加适量的甘油能获得较高的复性收率。

This article reports oxidative protein refolding assisted by artificial chaperones in reverse micelles formed by nonionic surfactant of sorbitan trioleate modified with Cibacron Blue F-3GA in n-hexane. The denatured/reduced lysozymewas used as a model protein and cetyltrimethylammonium bromide (CTAB) and-cyclodextrin as the artificial chaperones. The use of the artificial chaperones has proved to increase the refolding efficiency of denatured-reduced lysozyme at the concentration range studied (3.5-5.9 mg/mL). Moreover, the artificial chaperones increased the refolding yield in a wide range of urea concentrations. However, the optimal urea concentration range was little affected by the presence of the artificial chaperones.

研究了流加操作与稀释添加耦合辅助高浓度变性还原溶菌酶的复性，建立了相关的过程动力学模型。利用三态复性的过程模型对实验数据进行了拟合，获得了较好的拟合效果。利用模型分析了复性过程的动力学特性，结果表明，两者耦合可在较低的盐酸胍浓度（ mol·L-1）存在下使较高浓度的（5mg·ml-1）变性溶菌酶获得80%以上的复性收率；并且发现添加剂乙酰胺的辅助复性作用与盐酸胍相同，降低盐酸胍浓度，适当提高乙酰胺浓度即可使聚集体生成速率常数最小，酶的复性收率最大；但甘油与盐酸胍具有协同作用，只有在适当的盐酸胍浓度下添加甘油才可获得理想的复性效果。

为了促使多肽亲和配基在实际分离研究中得到广泛应用，利用Fmoc固相合成法对从噬菌体展示肽库中筛选得到的与胰岛素具有高特异性亲和的七肽片段（HWWWPAS）进行了合成。在合成中发现，肽段序列中脯氨酸的存在会严重影响合成结果。云除脯氨酸后，目标产物的收率从0%提高到11.6%；同时，七肽序列中丙氨酸的存在也会影响合成效果。若将丙氨酸除去，合成收率会明显提高（达到76%），特别是将丙氨酸更换为谷氨酸产物的收率和线度可达到70%以上。利用固定化六肽片段（HWWWES）进行亲和吸附实验的结果表明，改造后的六肽配基与原先的七肽配基具有相似的亲和吸附效果。